Adventitious Shoot Regeneration and Rooting of Prunus serotina In Vitro Cultures

Abstract

<jats:p>A complete regeneration protocol was developed for <jats:italic>Prunus serotina</jats:italic> Ehrh., an important hardwood species for timber and sawlog production in the central and eastern United States. Nodal sections were cultured on Murashige and Skoog (MS) medium supplemented with 4.44 μ<jats:sc>m</jats:sc> 6-benzylaminopurine (BA), 0.49 μ<jats:sc>m</jats:sc> indole-3-butyric acid (IBA), and 0.29 μ<jats:sc>m</jats:sc> gibberellic acid (GA<jats:sub>3</jats:sub>). In vitro leaf explants of three genotypes were placed on woody plant medium (WPM) supplemented with 0, 2.27, 4.54, or 6.81 μ<jats:sc>m</jats:sc> thidiazuron (TDZ) in combination with 0, 0.54, 1.07, or 5.37 μ<jats:sc>m</jats:sc> naphthaleneacetic acid (NAA), and on WPM supplemented with 0, 4.44, 8.88, or 13.32 μ<jats:sc>m</jats:sc> BA in combination with 0, 0.54, 1.07, or 5.37 μ<jats:sc>m</jats:sc> NAA. Cultures were maintained either in continuous darkness for 5 weeks, or in the dark for 3 weeks and then transferred to a 16-hour photoperiod. TDZ and the genotype had a significant effect on the number of shoots regenerated. The maximum mean number of shoots regenerated per explant (5.05 ± 1.14) was obtained with 2.27 μ<jats:sc>m</jats:sc> TDZ plus 0.54 μ<jats:sc>m</jats:sc> NAA with the 3-week dark period then light treatment. The highest percent shoot regeneration (38.3) and mean number of shoots (4.13 ± 0.97) was obtained with 6.81 μ<jats:sc>m</jats:sc> TDZ plus 1.07 μ<jats:sc>m</jats:sc> NAA. The highest rooting (27%) of adventitious shoots and number of roots per shoot (2.3 ± 0.2) was obtained with 2.5 μ<jats:sc>m</jats:sc> IBA when shoots were maintained for 7 days in the dark on rooting medium before transfer to a 16-hour photoperiod. The highest rooting (70%) of nodal explant-derived stock cultures and number of roots per shoot (2.7 ± 0.9) was also obtained with 2.5 μ<jats:sc>m</jats:sc> IBA, but when shoots were maintained for 4 days in the dark before transfer to a 16-hour photoperiod. In total, 86% of the plantlets survived acclimatization to the greenhouse and 100% survival after overwintering in cold-storage.</jats:p>

In vitro culture of Capparis decidua and assessment of clonal fidelity of the regenerated plants

“…1C). Similar results have been reported in other plants (Gyves et al 2001, Espinosa et al 2006. The interaction of cytokinins with low concentration of auxin enhancing shoot multiplication has been reported by many workers (Sharma et al 2003, Joshi and Kothari 2007, Santombi and Sharma 2008.…”

Plant regeneration from in vitro leaves of mature black cherry (Prunus serotina)

“…Cultures were incubated at 25°C under a 16-h photoperiod provided by cool-white fluorescent lamps (100-140 lmol m -2 s -1 ). Explants were transferred to fresh medium every 3 weeks for multiplication of shoots (see Espinosa et al 2006 for details).…”

“…Adventitious shoot regeneration using leaf explants is established for several species of Prunus, such as P. canescens (Antonelli and Druart 1990), P. padus (Hammatt 1993), P. domestica Cossio 1991, 1994;Nowak et al 2004), P. persica (Declerck and Korban 1996;Gentile et al 2002;Scorza et al 1995), P. dulcis (Ainsley et al 2000;Miguel and Oliveira 1999;Tang et al 2002), P. armeniaca (Perez-Tornero et al 2000), P. avium (Bhagwat and Lane 2004;Matt and Jehle 2005), and P. serotina (Espinosa et al 2006;Hammatt and Grant 1998).…”

“…A combination of cytokinin and auxin was also important for production of shoots from leaves. Thidiazuron (TDZ) produced a better response than 6-benzylaminopurine (BA) when inducing shoots from leaves of P. avium, P. armeniaca, and P. serotina (Espinosa et al 2006;Hammatt and Grant 1998;Perez-Tornero et al 2000). In P. avium, regeneration was optimal when whole-leaf explants were wounded and cultured abaxial side up, on medium supplemented with TDZ and napthaleneacetic acid (NAA) (Bhagwat and Lane 2004).…”

“…Hammatt and Grant (1998) were the first to report adventitious shoot regeneration from juvenile leaf explants of P. serotina, however, a complete protocol including rooting and acclimatization of regenerated plantlets was not established. Espinosa et al (2006) established a complete protocol for regeneration of plants of P. serotina from juvenile explants. The only report of Agrobacterium infection of black cherry was by Maynard and Fuernkranz (1989) using leaf pieces for regeneration and five Agrobacterium strains for gall formation tests.…”